Journal: BMC Cancer

Article Title: Reduced repair of 8-hydroxyguanine in the human breast cancer cell line, HCC1937

doi: 10.1186/1471-2407-6-297

Figure Lengend Snippet: Expression of antioxidants in HCC1937 and MCF-7 relative to AG11134 . Western Blot analysis of SOD1, SOD2 and catalase showing their levels in the two breast cancer cell lines compared to the normal mammary epithelial cell line, AG11134. β-actin was used as a loading control. The gels are representatives of three independent analyses in each case.

Article Snippet: HCC1937 is a primary ductal breast carcinoma cell line that was purchased from American Type Culture Collection (ATCC Manassas, VA).

Techniques: Expressing, Western Blot

Journal: NPJ Precision Oncology

Article Title: Interferon-γ signaling is associated with BRCA1 loss-of-function mutations in high grade serous ovarian cancer

doi: 10.1038/s41698-019-0103-4

Figure Lengend Snippet: Gene expression changes associated with BRCA1 mutation in ovarian cancer (OC) cells. a Survival (means ± s.e.m., n = 4) measured by the CCK8 assay of UWB1.289 OC cells (BRCA1-null) and UWB1.289 cells transfected with BRCA1 (BRCA1+) and treated with entinostat for 72 h. * P < 0.05 ( t -test). b , c Western blot and densitometric analysis (means ± s.e.m., n = 3) of entinostat effects (72 h treatment) on H3K9ac b and H3K27ac c levels in BRCA1-null and BRCA1+ UWB1.289 OC cells. * P < 0.05 ( t -test) relative to 0 entinostat. M, protein markers. d – f Unsupervised hierarchical clustering of differentially expressed genes measured by RNAseq between BRCA1-null and BRCA1+ d , entinostat-treated BRCA1+ compared with BRCA1+ e , and entinostat-treated BRCA1-null compared with BRCA1-null f UWB1.289 OC cells. Colored horizontal lines represent genes and columns represent specimens analyzed. Cells were treated with 0.5 μM entinostat for 24 h ( n = 2). g Numbers of differentially expressed genes in BRCA1-null relative to BRCA1+UWB1.289 OC cells treated with 0.5 μM entinostat for 24 h ( n = 2). h A Venn diagram shows numbers of common and unique differentially expressed genes determined by RNAseq in BRCA1+ and BRCA1-null UWB1.289 OC cells treated with 0.5 μM entinostat (Ent) for 24 h ( n = 2).

Article Snippet: UWB1.289 (BRCA1-null), UWB1.289+ BRCA1, and HCC1937 (BRCA1-null) cells lines were obtained authenticated and mycoplasma-free from the ATCC in 2017.

Techniques: Expressing, Mutagenesis, CCK-8 Assay, Transfection, Western Blot

Journal: NPJ Precision Oncology

Article Title: Interferon-γ signaling is associated with BRCA1 loss-of-function mutations in high grade serous ovarian cancer

doi: 10.1038/s41698-019-0103-4

Figure Lengend Snippet: Gene expression profiles associated with BRCA1 expression levels in ovarian cancer (OC) tumors. a Distribution of BRCA1 mRNA expression levels in high-grade serous ovarian cancer (HGSOC) specimens from the TCGA dataset. The upper 25% quantile (blue line to the right) were considered BRCA1-normal while the low 10% quantile (red line to the left) were considered BRCA1-deficient. b Volcano plot of differentially expressed genes between BRCA1-deficient and BRCA1-normal HGSOC tumors in the TCGA database. c Venn diagram shows the numbers of specific or shared genes differentially expressed between BRCA1-deficient and BRCA1-normal OC tumors (TCGA) and between BRCA1-null and BRCA1+ UWB1.289 OC cells. d IPA comparative analysis shows activation Z -scores of differentially expressed upstream regulators in BRCA1-null relative to BRCA1+ UWB1.289 OC cells, and in the same cell lines in response to entinostat treatment (0.5 μM for 24 h, n = 2).

Article Snippet: UWB1.289 (BRCA1-null), UWB1.289+ BRCA1, and HCC1937 (BRCA1-null) cells lines were obtained authenticated and mycoplasma-free from the ATCC in 2017.

Techniques: Expressing, Activation Assay

Journal: NPJ Precision Oncology

Article Title: Interferon-γ signaling is associated with BRCA1 loss-of-function mutations in high grade serous ovarian cancer

doi: 10.1038/s41698-019-0103-4

Figure Lengend Snippet: Top molecular and cellular functions identified by IPA among genes differing in expression (RNAseq, left) between BRCA1-null and BRCA1+ ovarian cancer (OC) cells, and among differentially expressed genes between BRCA1-deficient and BRCA1-normal OC tumors from the TCGA database (right).

Article Snippet: UWB1.289 (BRCA1-null), UWB1.289+ BRCA1, and HCC1937 (BRCA1-null) cells lines were obtained authenticated and mycoplasma-free from the ATCC in 2017.

Techniques: Expressing, Cell Function Assay

Journal: NPJ Precision Oncology

Article Title: Interferon-γ signaling is associated with BRCA1 loss-of-function mutations in high grade serous ovarian cancer

doi: 10.1038/s41698-019-0103-4

Figure Lengend Snippet: Top upstream regulators determined by IPA among genes differing in expression between BRCA1-null and BRCA1+ OC cells (left), and among differentially expressed genes between BRCA1-deficient and BRCA1-normal OC tumors from the TCGA database (right).

Article Snippet: UWB1.289 (BRCA1-null), UWB1.289+ BRCA1, and HCC1937 (BRCA1-null) cells lines were obtained authenticated and mycoplasma-free from the ATCC in 2017.

Techniques: Expressing, Activation Assay

Journal: NPJ Precision Oncology

Article Title: Interferon-γ signaling is associated with BRCA1 loss-of-function mutations in high grade serous ovarian cancer

doi: 10.1038/s41698-019-0103-4

Figure Lengend Snippet: Expression of IFN-regulated genes in BRCA1-null and BRCA1+ cancer cells. a mRNA expression levels (means ± s.e.m., n = 3) measured by qRT-PCR of the IFN-γ-regulated genes CXCL10 , CXCL11 , and IFI16 in BRCA1-null UWB1.289 OC cells with BRCA1 restored (BRCA1+) or transfected with empty vector (BRCA1-null). * P < 0.01 ( t -test). b Western blot measures STAT1 and p-STAT1 in BRCA1+ and BRCA1-null UWB1.289 OC cells treated with IFN-γ for 15 min. M, protein markers. c Expression levels (means ± s.e.m., n = 3) of the IFN-γ-regulated genes CXCL10, CXCL11 , and IFI16 in BRCA1+ and BRCA1-null OC cells treated with 10 ng/mL IFN-γ for 24 h. * P < 0.05 ( t -test) relative to control (0 dose) or between cell lines as indicated. d mRNA expression levels (means ± s.e.m., n = 3) of the IFN-α-regulated genes IFITM1 and MX1 in BRCA1+ and BRCA1-null UWB1.289 OC cells. * P < 0.05 ( t -test). e Measurements by western blot of STAT1 and p-STAT1 in BRCA1+ and BRCA1-null UWB1.289 OC cells treated with IFN-α for 15 min at the indicated doses. f Expression levels (means ± s.e.m., n = 3) of the IFN-α-regulated genes MX1 and IFITM1 in BRCA1+ and BRCA1-null UWB1.289 OC cells treated with 10 ng/mL IFN-α for 24 h. * P < 0.05 ( t -test) relative to control (0 dose) or between cell lines as indicated. g Binding levels (means ± s.e.m., n = 3) of STAT1 to the promoters of IFN-γ-regulated genes CXCL10 , CXCL11 , and IFI16 measured by ChIP in BRCA1+ and BRCA1-null UWB1.289 OC cells. * P < 0.05 ( t -test); F1 forward primer; R1 reverse primer.

Article Snippet: UWB1.289 (BRCA1-null), UWB1.289+ BRCA1, and HCC1937 (BRCA1-null) cells lines were obtained authenticated and mycoplasma-free from the ATCC in 2017.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Plasmid Preparation, Western Blot, Binding Assay

Journal: NPJ Precision Oncology

Article Title: Interferon-γ signaling is associated with BRCA1 loss-of-function mutations in high grade serous ovarian cancer

doi: 10.1038/s41698-019-0103-4

Figure Lengend Snippet: Cellular responses to IFN-γ depend upon BRCA1 mutational status. a Expression levels (means ± s.e.m., n = 3) of the CXCL10, CXCL11 , and IFI16 genes measured by qRT-PCR in HCC1937 (BRCA1-null) and HCC1937+ BRCA1 breast cancer (BC) cells treated with IFN-γ for 24 h. * P < 0.05 ( t -test) relative to control (0 h) or between cell lines as indicated. b CXCL10 , CXCL11 , and IFI16 mRNA expression levels in BRCA1-normal compared with BRCA1-mutated immortalized mammary epithelial cells. * P < 0.05 (mean ± s.e.m. of three independent measurements in two BRCA1-normal and one BRCA1-mutated cell cultures, t -test). c – f Cell survival (mean ± SE, n = 4) measured by the CCK8 assay of BRCA1-null and BRCA1+ ovarian cancer (OC) or BC cells treated with IFN-γ ( c , d ) or with IFN-α ( e , f ) for 96 h. * P < 0.05 ( t -test) between groups at the indicated doses. g STAT1 mRNA levels (means ± s.e.m., n = 2) measured by qRT-PCR in BRCA1+ and BRCA1-null UWB1.289 OC cells transduced with a non-targeting shRNA (shControl) or two different shRNAs directed at STAT1 . h CCK8 assay measured proliferation of BRCA1+ and BRCA1-null UWB1.289 OC cells transduced with shControl or shRNAs targeting STAT1 . NS not different ( P > 0.05, t -test). Values are percentage of cells (means ± s.e.m., n = 4) relative to day 1. Shown are t -test P values. i Effects of entinostat (Ent, 0.5 μM), IFN-γ (10 ng/mL), or combination treatment for 24 h on mRNA expression levels (means ± s.e.m., n = 3) of the CXCL10, CXCL11 , and IFI16 genes in BRCA1+ and BRCA1-null UWB1.289 OC cells. * P < 0.05 ( t -test) relative to Control or between BRCA1+ treated with IFN-γ vs. Ent+ IFN-γ for IFI16 .

Article Snippet: UWB1.289 (BRCA1-null), UWB1.289+ BRCA1, and HCC1937 (BRCA1-null) cells lines were obtained authenticated and mycoplasma-free from the ATCC in 2017.

Techniques: Expressing, Quantitative RT-PCR, CCK-8 Assay, Transduction, shRNA

Journal: NPJ Precision Oncology

Article Title: Interferon-γ signaling is associated with BRCA1 loss-of-function mutations in high grade serous ovarian cancer

doi: 10.1038/s41698-019-0103-4

Figure Lengend Snippet: Differential histone H3K9 and H3K27 acetylation in ovarian cancer (OC) cells carrying a BRCA1 loss-of-function mutation (BRCA1-null). a Venn diagrams illustrate numbers of unique and common H3K9ac or H3K27ac peaks in BRCA1-null and BRCA1+ UWB1.289 OC cells measured by ChIP-seq. b Percent distribution of all H3K9ac or H3K27ac peaks among genomic regions in BRCA1+ and BRCA1-null UWB1.289 OC cells. c Percent distribution among genomic regions of differential H3K9ac or H3K27ac peaks in BRCA1-null vs. BRCA1+ UWB1.289 OC cells. d Differential H3K27ac peaks (−5000 bp to +5000 bp from the peak center) grouped into four clusters and represented as integrated plots (top) or a heatmap (bottom) in BRCA1-null vs. BRCA1+ UWB1.289 OC cells. Rows on the heatmap represent H3K27ac peaks. e Venn diagrams shows numbers of common genes having differential promoter H3K9ac or H3K27ac marks (ChIP-seq, blue) and differential expression (RNAseq, red) between BRCA1-null and BRCA1+ UWB1.289 OC cells. f Profiles of H3K9ac and H3K27ac peaks in the IFN-γ-regulated gene IFI16 in BRCA1-null vs. BRCA1+ UWB1.289 OC cells. g H3K9ac enrichment (means ± s.e.m., n = 2) measured by ChIP-PCR in the promoters of the IFN-γ-regulated genes CXCL10 , CXCL11 , and IFI16 in BRCA1-null and BRCA1+ UWB1.289 OC cell line * P < 0.05 ( t -test).

Article Snippet: UWB1.289 (BRCA1-null), UWB1.289+ BRCA1, and HCC1937 (BRCA1-null) cells lines were obtained authenticated and mycoplasma-free from the ATCC in 2017.

Techniques: Mutagenesis, ChIP-sequencing, Expressing

Journal: Acta Histochemica et Cytochemica

Article Title: Stem-like Human Breast Cancer Cells Initiate Vasculogenic Mimicry on Matrigel

doi: 10.1267/ahc.18041

Figure Lengend Snippet: VM formation of HCC1937/p53 cells on matrigel. a ) b ) Microscopic appearances of the cellular structures 24 hr after seeding on matrigel are photographed using an inverted phase contrast microscope. a : Cultured in flask for 24 hr. b : Cultured on matrigel (Bar = 200 μm). The aggregated regions (asterisks) are 4 or more cells thick, whereas the bridging regions (arrowheads) are 1–3 cells thick. c ) d ) The cells were stained with DAPI and photographed using a confocal laser scanning microscope. The 3D structures in the B regions are shown using Temporal-Color Code of Fiji/ImageJ software. Twenty-four vertical serial sections (each 1.0 μm thick) were stacked and displayed with the color code according to the depth of the nuclear location. c : Cells cultured on APS-coated slides for 24 hr. d : Cells cultured on matrigel for 24 hr (Bar = 100 μm).

Article Snippet: The human breast cancer cell line HCC1937 was purchased from American Type Culture Collection (ATCC).

Techniques: Microscopy, Cell Culture, Staining, Laser-Scanning Microscopy, Software

Journal: Acta Histochemica et Cytochemica

Article Title: Stem-like Human Breast Cancer Cells Initiate Vasculogenic Mimicry on Matrigel

doi: 10.1267/ahc.18041

Figure Lengend Snippet: The expression patterns of ALDH1A3 and Sox-2 in Dox-treated cells. The time-course changes of the percentage of ALDH1A3 + cells and Sox-2 + cells in our previous study are shown. Residual HCC1937/p53 cells indicate apoptosis-resistant cells in those showing Dox-induced p53 overexpression .

Article Snippet: The human breast cancer cell line HCC1937 was purchased from American Type Culture Collection (ATCC).

Techniques: Expressing, Over Expression

Journal: Acta Histochemica et Cytochemica

Article Title: Stem-like Human Breast Cancer Cells Initiate Vasculogenic Mimicry on Matrigel

doi: 10.1267/ahc.18041

Figure Lengend Snippet: Accelerated VM formation on matrigel in apoptosis-resistant subpopulations of HCC1937/p53 cells. A : The cells on doxycycline for 2 days were seeded in matrigel and cultured. The control cells were not treated with doxycycline. B : The time-course changes of VM scores were calculated by the formula in . Each line graph is modified by linear regression (Control: R 2 = 0.9927, Dox2d: R 2 = 0.9043) and shown as a dotted line.

Article Snippet: The human breast cancer cell line HCC1937 was purchased from American Type Culture Collection (ATCC).

Techniques: Cell Culture, Modification